大鼠早期局灶性脑缺血再灌注损伤中USP10的表达水平变化及其与自噬的关系  

The study on the relation between the expression of USP10 and autophagy following early focal cerebral ischemical reperfusion injury in rats

作　　者：方聪聪[1] 毛善平[1] 曾智[2] 董慧敏[1] 刘宝辉[3] 王舜[1] 谭华威 

机构地区：[1]武汉大学人民医院神经内科,430060 [2]武汉大学人民医院病理科,430060 [3]武汉大学人民医院神经外科,430060 [4]武汉大学人民医院精神科,430060

出　　处：《卒中与神经疾病》2017年第4期290-296,共7页Stroke and Nervous Diseases

基　　金：武汉市科技攻关计划项目(项目编号为2013060602010270)

摘　　要：目的探讨早期局灶性脑缺血再灌注损伤大鼠模型中再灌注不同时间点USP10的表达水平变化及其与自噬的关系。方法将36只成年雄性SD大鼠随机分成4组:假手术组、脑缺血2 h再灌注6 h模型组、脑缺血2 h再灌注12 h模型组、脑缺血2 h再灌注24 h模型组;采用线栓法致大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)制备大鼠局灶性脑缺血再灌注模型,给予神经行为学评分,用TTC染色法测定脑梗死体积,透射电镜观察梗死周边区皮层神经元自噬,Western blot法检测梗死周边区皮层USP10和自噬相关蛋白LC3B的表达水平,免疫荧光双标法检测梗死周边区皮层神经元中USP10及LC3B的表达水平变化。结果免疫荧光双标显示模型组自噬蛋白阳性细胞数与存活神经元数均于再灌注12 h最多(P<0.05),二者某种程度上趋势一致;Western blot免疫荧光双标均显示与假手术组相比,USP10及LC3B蛋白在模型组中表达上调(P<0.01),且于再灌注12 h组达到高峰(P<0.05);透射电镜显示再灌注不同时间点自噬强度的变化与免疫荧光自噬蛋白表达水平的趋势基本一致。结论早期脑I/R损伤中自噬的激活某种程度上可减轻神经元的损伤,而USP10可能通过某种机制调控脑I/R中自噬的发生发展。Objective To explore the relation between USP10 expression and autophagy in rats model of early focal cerebral ischemia-reperfusion（I/R） injury.Methods Totally 36 male,healthy Sprague-Dawley rats were used and randomlyassigned to four groups： Sham-operated group（Sham） and cerebral ischemia-reperfusion group （MCAO6 h,12 h and 24 h group）.The focal cerebral chemia-reperfusiong （I/R） rat models were established by the middle cerebral artery occlusion （MCAO） using intraluminal suture method.The behavior scales function rats was used as a general assessment of neural of brain injury;2,3,5-triphenyltetrazolium chloride （TTC） straining was used to observe infarct volume;specific structure of autophagosome and specific protein of autophagy microtubule-associated protein 1 light chain 3 B（LC3B）were detected by transmission electron microscope,WB and immunofluorescence respectively.Results Compared to the Sham group,autophagy existed in different time periods after I/R shown both in transmission electron microscope,WB and immunofluorescence,which was in line with the expression of USP10.The number of positive cells and survival neurons was significantly increased at I/R 12H compared with sham group（P〈0.05） in immunofluorescence.Conclusion The intensity of autophagy was positive correlation of the number of survival neurons in rats model of early focal cerebral ischemia-reperfusion（I/R） injury.The expression of USP10 was in accord with LC3B,which indicates USP10 might regulate the development of autophagy and would lay important foundation for the study on the interaction of autophagy-lysosomes system and ubiquitin-proteasomes system in cerebral I/R injury.

关 键 词：脑缺血再灌注损伤 自噬 USP10 LC3 

分 类 号：R743.33[医药卫生—神经病学与精神病学]